U.S. patent application number 10/268180 was filed with the patent office on 2003-05-08 for method for inhibiting the formation of seromas using factor xiii.
Invention is credited to Bishop, Paul D., Grossmann, Angelika.
Application Number | 20030087825 10/268180 |
Document ID | / |
Family ID | 23279383 |
Filed Date | 2003-05-08 |
United States Patent
Application |
20030087825 |
Kind Code |
A1 |
Bishop, Paul D. ; et
al. |
May 8, 2003 |
Method for inhibiting the formation of seromas using factor
XIII
Abstract
Use of factor XIII for inhibiting the formation of seromas by
administering factor XIII. The factor XIII can be administered
locally at the site of a wound or surgery or administered
systemically. If the factor XIII is administered locally, it can be
activated or non-activated and may be administered in conjunction
with activated thrombin.
Inventors: |
Bishop, Paul D.; (Fall City,
WA) ; Grossmann, Angelika; (Seattle, WA) |
Correspondence
Address: |
Paul G. Lunn, Esq.
ZymoGenetics, Inc.
1201 Eastlake Avenue East
Seattle
WA
98102
US
|
Family ID: |
23279383 |
Appl. No.: |
10/268180 |
Filed: |
October 9, 2002 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
60328070 |
Oct 9, 2001 |
|
|
|
Current U.S.
Class: |
424/94.64 |
Current CPC
Class: |
A61K 2300/00 20130101;
A61P 7/00 20180101; A61K 38/45 20130101; A61P 17/02 20180101; A61K
38/4833 20130101; A61K 38/4833 20130101 |
Class at
Publication: |
514/12 |
International
Class: |
A61K 038/37 |
Claims
What is claimed is:
1. A method for inhibiting the formation of a seromal or serous
fluid at the site of surgery or a wound in a mammal comprising
administering to said mammal a therapeutically effective amount of
factor XIII.
2. The method of claim 1 wherein the factor XIII is administered
systemically.
3. The method of claim 1 wherein the factor XIII is administered
locally at the site of the surgery or wound.
4. The method of claim 3 wherein the factor XIII is activated.
5. The method of claim 3 wherein the factor XIII is administered in
conjunction with activated thrombin.
Description
[0001] This claims priority under 35 U.S.C. .sctn.119(e) of U.S.
Provisional Application No. 60/328,070 filed Oct. 9, 2001.
BACKGROUND OF THE INVENTION
[0002] Seromas are collections of lymph usually present as painless
swellings within a wound or below flaps. These often develop in
wounds involving dissection in lymph node-bearing areas, for
example axillae, neck, groin etc., or in areas where significant
dead space remains such as after abdominal-perineal resection,
total mastectomy or in breast reduction procedures either for
females or to treat gynecomastia in males. The seromas prevent
adequate tissue approximation or may become secondarily
infected.
[0003] The primary cause lies in the failure to identify and
control lymphatic vessels during dissection. Though lymph is a
protein-rich fluid, electrocauterization is ineffective to prevent
seroma formation. Thus, there is a need to develop a treatment to
prevent the formation of seromas.
DESCRIPTION OF THE INVENTION
[0004] The present invention fills this need by administering
factor XIII to patients who have undergone surgery to inhibit the
build-up of fluids or seromas beneath the skin where the surgery
took place or a wound has occurred. The factor XIII may be applied
locally in solution beneath the skin or administered systemically.
The factor XIII solution can be administered prior to surgery,
prior to suturing of the surgical site or can be injected beneath
the skin after surgery. If the factor XIII is administered locally,
it may be activated or non-activated, or the non-activated factor
XIII may be applied in conjunction with activated alpha-thrombin.
The activated thrombin would then activate the factor XIII.
Activated thrombin can be administered locally at a concentration
of about 0.5 mg/mL of solution. A method for producing human
recombinant thrombin can be found in U.S. Pat. No. 5,502,034.
locally at a concentration of about 0.5 mg/mL of solution. A method
for producing human recombinant thrombin can be found in U.S. Pat.
No. 5,502,034.
[0005] Factor XIII, also known as fibrin-stabilizing factor,
circulates in the plasma at a concentration of 20 .mu.g/ml. The
protein exists in plasma as a tetramer comprised of two A subunits
and two B subunits. Each subunit has a molecular weight of 83,000
Da, and the complete protein has a molecular weight of
approximately 330,000 Da. Factor XIII catalyzes the cross-linkage
between the .gamma.-glutamyl and .epsilon.-lysyl groups of
different fibrin strands. The catalytic activity of factor XIII
resides in the A subunits. The B subunits act as carriers for the A
subunits in plasma factor XIII.
[0006] Recombinant factor XIII can be produced according to the
process described in European Patent No. 0 268 772 B1. The level of
factor XIII in the plasma can be increased by administering a
factor XIII concentrate derived from human placenta called
FIBROGAMMIN.RTM. (Aventis Corp.) or by administration of
recombinant factor XIII.
[0007] As stated above, administration of factor XIII to a subject
is may be administered locally at the site of the wound or
systemically. If administered systemically, the factor XIII is
generally administered intravenously. When administering
therapeutic proteins by injection, the administration may be by
continuous infusion or by single or multiple boluses. A
pharmaceutical composition comprising factor XIII can be formulated
according to known methods to prepare pharmaceutically useful
compositions, whereby the therapeutic proteins are combined in a
mixture with a pharmaceutically acceptable carrier. A composition
is said to be a "pharmaceutically acceptable carrier" if its
administration can be tolerated by a recipient patient. A suitable
pharmaceutical composition of factor XIII will contain 1 mM EDTA,
10 mM glycine, 2% sucrose in water. An alternative formulation will
be a factor XIII composition containing 20 mM histidine, 3%
wt/volume sucrose, 2 mM glycine and 0.01% wt/vol. polysorbate, pH
8.
[0008] Other suitable carriers are well known to those in the art.
See, for example, Gennaro (ed.), Remington's Pharmaceutical
Sciences, 19th Edition (Mack Publishing Company 1995).
[0009] Administration of Factor XIII
[0010] The levels of factor XIII in an individual can be determined
by assays well known in the art such as the BERICHROM.RTM.F XIII
assay (Dade Behring Marburgh GmbH, Marburg, Germany). The normal
adult has an average of about 45 ml of plasma per kg of body
weight. Each liter of blood has 1000 units (U) of factor XIII. The
amount of factor XIII administered should be enough to bring an
individual's level of factor XIII in the plasma to 100% of normal
plasma or slightly above to 1-5% above normal. A dose of 0.45 U/kg
would raise the level of factor XIII by about 1% compared to
normal. One unit of factor XIII is about 10 .mu.g of recombinant
factor XIII, which contains only the dimerized A subunit. Thus, to
raise the level of factor XIII by 1%, one would administer about
4.5 .mu.g of the A2 subunit per kilogram weight of the individual.
So to raise the level 30% of normal, one would administer 13.5
U/kg. For a 75 kg individual this would be about 1,012.5 U. Some
patients may have consumptive coagulopathies that involve factor
XIII losses. In such cases, a higher dosing (e.g., 1-2 U/kg-%) or
multiple dosing of factor XIII (e.g., 1-2 U/kg-%-day) may be
required.
EXAMPLE 1
The Use of Factor XIII to Prevent Seroma Formation in a Rat Seromal
Mastectomy Model
[0011] Object of the Experiment
[0012] The object of the experiment was to determine if factor XIII
when given systemically would influence seromal fluid formation
using a rat mastectomy model.
[0013] Background
[0014] Seromas are the most common postoperative complication for
patients undergoing a mastectomy. The formation of these fluid
collections is facilitated by the disruption of lymphatics and
blood vessels as well as by the creation of large potential voids
beneath the skin. Postoperative problems due to seroma formation
include delayed wound healing, flap necrosis, lymph edema of the
arm and infection.
[0015] Factor XIII PreparationFactor XIII was provided by
ZymoGenetics, Inc., Seattle Wash. in bottles containing 13.2 mg of
lyophilized factor XIII containing 0.3 mM
ethylenediaminetetraacetic acid (EDTA), 31 mM glycine, and 6.2%
sucrose. The lyophilized factor XIII was reconstituted with 3.3 mL
of sterile water and pipetted into 0.5 mL aliquots and stored in a
freezer at -20.degree. C.
[0016] Vehicle Preparation
[0017] The vehicle preparation was a lyophilized powder comprised
of 0.3 mM EDTA, 31 mM glycine and 6.2% sucrose. This was
reconstituted with 3.3 ml of sterile water and pipetted into 0.5 mL
aliquots and stored in a freezer at -20.degree. C. After thawing,
0.5 mL of bovine serum albumin (BSA) was added to each vial.
[0018] Experimental Procedure
[0019] The rats were anesthetized with isoflurane (3% isoflurane,
1% oxygen) and a catheter was inserted into the jugular vein of
each rat. Two to three days following catheter insertion, eleven
rats received a single intravenous bolus injection through the
catheter of the vehicle preparation (the control group) and 12 rats
received a single bolus injection of 1 mg/kg of the factor XIII
preparation (the experimental group) 30 minutes prior to a left
side radical mastectomy. The mastectomy consisted of removal of the
pectoralis muscle, lymphatic tissue including nodes (3 or more) and
traumatization of subcutaneous lymphovasculature surface. The
traumatization the lymphovasculature occurred by scraping 50 times
the inner surface of the elevated skin flap with a No. 22 scalpel
blade. Five days following mastectomy, the rats were anesthetized
with urethane anesthesia. Blood samples were taken for analysis.
Seromal fluid was aspirated and weighed from each rat to determine
seromal fluid volume and factor XIII content.
[0020] Results
[0021] No significant differences were observed between the Control
and Experimental rats for the following: hematology, clinical blood
chemistry and factor XIII levels.
[0022] A significant difference was observed in seromal fluid
volume aspirate at time of sacrifice (day 5). The control group
averaged 2.7 mL of seromal fluid and the group to whom factor XIII
was administered averaged 1.4 mL of seromal fluid. Thus factor XIII
was effective in inhibiting the formation of seromal fluid.
* * * * *